Abadan Faculty of Medical Science, Abadan, Iran.
Cellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
Acta Histochem. 2022 Aug;124(6):151928. doi: 10.1016/j.acthis.2022.151928. Epub 2022 Jul 4.
Diabetes is a chronic disease that is associated with a decrease or disfunction of β-cell. In the present study, fabrication of bioartificial pancreas using MIN-6 β-cell line seeded in decellularized rat testicles was investigated.
In this experimental study, the whole body of testes were decellularized and after characterization, were seeded by MIN-6 cell line. The expression of insulin-related genes and proteins including PDX-1, Glut2, Insulin, and Neurogenin-3 were evaluated. Insulin secretion was assessed under different concentrations of glucose. Seeded scaffolds with or without MIN-6 cells were transplanted to the rat's mesentery and their blood sugar and body weight were evaluated every three days for 28 days and analyzed with H&E staining.
Histological assessments indicated the cells were completely removed after decellularization. The scaffold had no toxic impacts on the MIN-6 cells (P˂ 0.02). Insulin release in response to different concentrations of glucose in 3D culture (testis-ECM) was significantly more than the traditional 2D monolayer culture (P < 0.001). Moreover, the relative genes and proteins expression were significantly higher in the 3D culture, compared to the 2D control group. In vivo transplantation of the (testis- Extra Cellular Matrix) testis-ECM scaffolds showed appropriate positions for transplantation with angiogenesis and low infiltration of inflammatory cells. The recellularized scaffolds could drop blood sugar levels and increase the body-weight of STZ-diabetic rats (P < 0.01).
Our study clearly confirmed that ECM valuable organ scaffolds prepared by decellularization of the testicular tissue is suitable for the fabrication of bioartificial pancreas for transplantation.
糖尿病是一种慢性疾病，与β细胞减少或功能障碍有关。本研究旨在探讨利用种植于去细胞化大鼠睾丸的 MIN-6β细胞系构建生物人工胰腺。
在这项实验研究中，对睾丸进行了整体去细胞化，然后对 MIN-6 细胞系进行了种植。评估了与胰岛素相关的基因和蛋白质的表达，包括 PDX-1、Glut2、胰岛素和 Neurogenin-3。评估了在不同葡萄糖浓度下的胰岛素分泌情况。将种植有或没有 MIN-6 细胞的支架移植到大鼠的肠系膜中，每隔三天评估其血糖和体重，并进行 H&E 染色分析。
组织学评估表明，去细胞化后细胞已完全去除。支架对 MIN-6 细胞没有毒性影响（P<0.02）。在 3D 培养（睾丸-细胞外基质）中，胰岛素对不同浓度葡萄糖的释放明显高于传统的 2D 单层培养（P<0.001）。此外，与 2D 对照组相比，3D 培养中的相对基因和蛋白质表达显著更高。体内移植（睾丸-细胞外基质）睾丸-ECM 支架显示出适当的移植位置，具有血管生成和低炎症细胞浸润。再细胞化的支架可以降低 STZ 糖尿病大鼠的血糖水平并增加体重（P<0.01）。
我们的研究清楚地证实，通过去细胞化睾丸组织制备的有价值的细胞外基质器官支架适合用于构建用于移植的生物人工胰腺。
